[关键词]
[摘要]
目的:对《中国药典》中记载的14种藤类药材进行分子鉴定。方法:以核基因ITS2片段作为DNA条形码,对研究材料进行PCR 扩增并双向测序,所得序列经CodonCode Aligner 拼接后,用软件MEGA4.0进行相关数据分析,并构建NJ(邻接)树。结果:14种藤类药材的ITS2 序列长度为190~284 bp;各药材种内K2P 遗传距离为0~0.053,远远小于种间K2P 遗传距离(平均值为0.852,最小值为0.309);由所构建的系统聚类树图可以看出,本研究中具有不同产地来源样品的药材均表现出了单系性,同时又与其它药材明显分开。结论:ITS2 序列作为条形码适用于鉴定《中国药典》中藤类药材,在中药材的鉴定领域具有重要的应用前景。
[Key word]
[Abstract]
This study was aimed to discriminate 14 types of vine herbs of the Chinese Pharmacopoeia. The second internal transcribed spacer (ITS2) of ribosomal DNA was amplified and sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. Phylogenetic study was performed using software MEGA 4.0 in accordance with the Kimura 2-Parameter (K2P) model. And the phylogenetic tree was constructed using neighbor-joining (NJ) methods. The results showed that the length of ITS2 sequence of 14 studied vine herbs was from 190 bp to 284 bp. The intraspecific genetic distance (K2P distance) of vine herbs is between 0 and 0.053, which was far lower than their interspecific genetic distance (mean: 0.852, the smallest: 0.309). In the cluster dendrogram, all studied medicinal materials with several samples showed monophyletic, and meanwhile, distinguished from others. It was concluded that ITS2 barcode is suitable for the identification of vine herbs from the Chinese Pharmacopoeia. This method will play an important role in the identification of Chinese Materia Medica.
[中图分类号]
[基金项目]
国家卫生部卫生行业科研专项(200802043):药用植物DNA barcoding(条形码)鉴定研究,负责人:陈士林;科学技术部国家科技支撑计划(2011BAI06B01):金银花规范化种植基地优化升级及系列产品综合开发研究,负责人:张永清;山东省科技厅科技发展计划(2008GG2NS02022):山东道地药材资源质量控制科技平台建设,负责人:张永清。